The aim of this study was to assess the clinical value of diagnosing and locating lower gastrointestinal (GI) bleeding using single photon emission computed tomography (SPECT)/computed tomography (CT) fusion imaging with 99mTc labeled red blood cells (99mTc-RBC). Fifty-six patients with suspected lower GI bleeding received a preoperative intravenous injection of 99mTc-RBC and each underwent planar, SPECT/CT imaging of the lower abdominal region. The location and path of lower GI bleeding were diagnosed by contrastive analysis of planar and SPECT/CT fusion imaging. Among the 56 patients selected, there were abnormalities in concentrated radionuclide activity with planar imaging in 50 patients and in SPECT/CT fusion imaging in 52 patients. Moreover, bleeding points that were coincident with the surgical results were evident with planar imaging in 31 patients and with SPECT/CT fusion imaging in 48 patients. The diagnostic sensitivity of planar imaging and SPECT/CT fusion imaging were 89.3% (50/56) and 92.9% (52/56), respectively, and the difference was not statistically significant (χ2 = 0.11, P > 0.05). The corresponding positional accuracy values were 73.8% (31/42) and 92.3% (48/52), and the difference was statistically significant (χ2 = 4.63, P 99mTc- RBC SPECT/CT fusion imaging is an effective, simple, and accurate method that can be used for diagnosing and locating lower GI bleeding.